Fig. 6
Mitochondrial calcium buffering inhibits ROS/Nrf2/Notch pathway and MCUR1-induced EMT and HCC metastasis. a Western blot analysis for expression levels of Nrf2, Notch1and NICD1 in stably transfected HCC cells with treatments as indicated. Mitochondrial Ca2+ was buffered by transient transfection of expression vector encoding parvalbumin with mitochondria target sequence (PV-Mito) for 48 h, where appropriate. b Western blot analysis for expression levels of ZO-1, E-cadherin, N-cadherin, Vimentin and Snail in stably transfected HCC cells with treatments as indicated. c Transwell assays for migration and invasion ability and wound healing assays for migration rate in stably transfected HCC cells with treatment as indicated. d Representative immunohistochemical staining images of E-cadherin and Vimentin in EV, MCUR1 and the Ad-PV-Mito groups (injection of PV-Mito adenovirus into tail vein). e Histological analyses of intrahepatic and lung metastatic nodules in EV, MCUR1 and the Ad-PV-Mito group (injection of PV-Mito adenovirus into tail vein). Images showing representative H&E staining of liver and lung tissue samples from the different experimental groups (n = 5 /group). f The number of intrahepatic and lung metastasis nodules was quantified in H.E. sections. g The schematic for mechanism underlying epithelial-mesenchymal transition and metastasis facilitated by MCUR1. Data shown are the mean ± SD from three independent experiments. * P < 0.05; ** P < 0.01